Translocation of human ribosomal protein S3 to sites of DNA damage is dependant on ERK-mediated phosphorylation following genotoxic stress

Yadavilli, Sridevi; Hegde, Vijay; Deutsch, Walter

doi:10.1016/j.dnarep.2007.04.009

Cited by 52 publications

(66 citation statements)

References 43 publications

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“…Our finding of a dose-dependent increase in nuclear localization of RPS3 under conditions of oxidative and UV stress (Fig. 8) is consistent with previous report (17). We did not observe discrete foci formation between RPS3 and RECQL4 possibly because of homogenous RECQL4 expression.…”

Section: Discussionsupporting

confidence: 82%

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Ribosomal Protein S3 Negatively Regulates Unwinding Activity of RecQ-like Helicase 4 through Their Physical Interaction

Patil

Hsieh

2017

Journal of Biological Chemistry

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Edited by Patrick SungHuman RecQ-like helicase 4 (RECQL4) plays crucial roles in replication initiation and DNA repair; however, the contextual regulation of its unwinding activity is not fully described. Mutations in RECQL4 have been linked to three diseases including Rothmund-Thomson syndrome, which is characterized by osteoskeletal deformities, photosensitivity, and increased osteosarcoma susceptibility. Understanding regulation of RECQL4 helicase activity by interaction partners will allow deciphering its role as an enzyme and a signaling cofactor in different cellular contexts. We became interested in studying the interaction of RECQL4 with ribosomal protein S3 (RPS3) because previous studies have shown that RPS3 activity is sometimes associated with phenotypes mimicking those of mutated RECQL4. RPS3 is a small ribosomal protein that also has extraribosomal functions, including apurnic-apyrimidinic endonuclease-like activity suggested to be important during DNA repair. Here, we report a functional and physical interaction between RPS3 and RECQL4 and show that this interaction may be enhanced during cellular stress. We show that RPS3 inhibits ATPase, DNA binding, and helicase activities of RECQL4 through their direct interaction. Further domain analysis shows that N-terminal 1-320 amino acids of RECQL4 directly interact with the C-terminal 94 -244 amino acids of RPS3 (C-RPS3). Biochemical analysis of C-RPS3 revealed that it comprises a standalone apurnic-apyrimidinic endonuclease-like domain. We used U2OS cells to show that oxidative stress and UV exposure could enhance the interaction between nuclear RPS3 and RECQL4. Regulation of RECQL4 biochemical activities by RPS3 along with nuclear interaction during UV and oxidative stress may serve to modulate active DNA repair.The RecQ class of helicases function at various stages of DNA metabolism. The presence of at least one homolog in each species evidences their crucial role in maintaining genomic stability. In humans, three of five RecQ helicases have been associated to different genetic disorders. WRN and BLM have been linked with Werner's syndrome and Bloom's syndrome, respectively, and RECQL4 has been linked with Rothmund-Thomson syndrome (RTS), 2 Baller-Gerald syndrome, and RAPADILINO syndrome. Of these, RECQL4 is unique on the basis of its domain structure and function. The N terminus of this protein represents the only human homolog of Saccharomyces cerevisiae essential replication initiation protein Sld2 (1, 2). The helicase domain is conserved across the RecQ class of helicases. Initial reports suggested RECQL4 lacked of helicase activity (3), but annealing, strand exchange and helicase activities of RECQL4 were later demonstrated (4 -7). Human RECQL4 was shown to unwind up to 22-bp double-stranded regions through its helicase activity (6, 7). ATPase and annealing activities of human RECQL4 have also been extensively studied (7,8). Robust helicase and annealing activities on substrates of various lengths have been reported for Drosophila melanogaster R...

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Section: Discussionsupporting

confidence: 82%

“…Ribosomal protein S3 (RPS3) is a component of the 40 S ribosomal subunit (16). However, nuclear localization of RPS3, with or without different stimulations (including hydrogen peroxide, methyl methane sulfonate, and TNF) has been reported (17)(18)(19). A mass spectrometry study revealed RPS3 as an interaction partner of WRN (20).…”

mentioning

confidence: 99%

Ribosomal Protein S3 Negatively Regulates Unwinding Activity of RecQ-like Helicase 4 through Their Physical Interaction

Patil

Hsieh

2017

Journal of Biological Chemistry

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“…In both yeast and humans, RpS3 has enzymatic activity on a variety of damaged DNA substrates generated by UV and oxidative stress (Jung et al 2001;Hegde et al 2004). In humans, RpS3 translocates to the nucleus from the cytosol after phosphorylation by ERK1/2 in cells exposed to oxidative stress and colocalizes with 8-oxoG foci in the nucleus (Yadavilli et al 2007). Titration of free RpS3 by overexpressed Ltv1DNES might deleteriously affect growth if free extraribosomal RpS3 plays a role in DNA surveillance and, hence, in cell division in yeast.…”

Section: Does Titration Of Rps3 Affect An Extraribosomal Function?mentioning

confidence: 99%

Genetic Analysis of the Ribosome Biogenesis Factor Ltv1 ofSaccharomyces cerevisiae

et al. 2014

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Ribosome biogenesis has been studied extensively in the yeast Saccharomyces cerevisiae. Yeast Ltv1 is a conserved 40S-associated biogenesis factor that has been proposed to function in small subunit nuclear export. Here we show that Ltv1 has a canonical leucine-rich nuclear export signal (NES) at its extreme C terminus that is both necessary for Crm1 interaction and Ltv1 export. The C terminus of Ltv1 can substitute for the NES in the 60S-export adapter Nmd3, demonstrating that it is a functional NES. Overexpression of an Ltv1 lacking its NES (Ltv1ΔC13) was strongly dominant negative and resulted in the nuclear accumulation of RpS3-GFP; however, export of the pre-40S was not affected. In addition, expression of endogenous levels of Ltv1ΔC protein complemented both the slow-growth phenotype and the 40S biogenesis defect of an ltv1 deletion mutant. Thus, if Ltv1 is a nuclear export adapter for the pre-40S subunit, its function must be fully redundant with additional export factors. The dominant negative phenotype of Ltv1ΔNES overexpression was suppressed by co-overexpressing RpS3 and its chaperone, Yar1, or by deletion of the RpS3-binding site in Ltv1ΔNES, suggesting that titration of RpS3 by Ltv1ΔNES is deleterious in yeast. The dominant-negative phenotype did not correlate with a decrease in 40S levels but rather with a reduction in the polysome-to-monosome ratio, indicating reduced rates of translation. We suggest that titration of RpS3 by excess nuclear Ltv1 interferes with 40S function or with a nonribosomal function of RpS3.

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“…The rpS3 protein is phosphorylated by extracellular signal-regulated kinase (ERK), a MAP kinase known to play important roles in the regulation of cell growth (6). The phosphorylation event mediated by ERK1/2 is necessary for rpS3 nuclear translocation in response to DNA damage (2). Kim et al have also reported that rpS3 has endonuclease activity, which is increased by protein kinase C delta (PKCδ)-dependent phosphorylation upon exposure to genotoxic stresses (3).…”

Section: Introductionmentioning

confidence: 97%

“…In addition, rpS3 has been shown to have additional extraribosomal functions, such as DNA repair, gene transcription, and apoptosis (2)(3)(4)(5). The rpS3 protein is phosphorylated by extracellular signal-regulated kinase (ERK), a MAP kinase known to play important roles in the regulation of cell growth (6).…”

Section: Introductionmentioning

confidence: 99%

Ribosomal protein S3 is phosphorylated by Cdk1/cdc2 during G2/M phase

Yoon¹,

Chung²,

Hahm³

et al. 2011

BMB Reports

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Translocation of human ribosomal protein S3 to sites of DNA damage is dependant on ERK-mediated phosphorylation following genotoxic stress

Cited by 52 publications

References 43 publications

Ribosomal Protein S3 Negatively Regulates Unwinding Activity of RecQ-like Helicase 4 through Their Physical Interaction

Ribosomal Protein S3 Negatively Regulates Unwinding Activity of RecQ-like Helicase 4 through Their Physical Interaction

Genetic Analysis of the Ribosome Biogenesis Factor Ltv1 ofSaccharomyces cerevisiae

Ribosomal protein S3 is phosphorylated by Cdk1/cdc2 during G2/M phase

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